1. Generation of a Universal Human Complement Source by Large-Scale Depletion of IgG and IgM from Pooled Human Plasma
- Author
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Holly E. Humphries, Lauren Allen, Frances Alexander, Andrew Gorringe, Rachel Halkerston, Elodie Lesne, Stephen Taylor, Emily Brunt, Stephen Thomas, Elizabeth J Penn, Breeze E. Cavell, and Stephanie Leung
- Subjects
Sepharose ,Immune system ,Affinity chromatography ,biology ,Immunity ,Chemistry ,biology.protein ,Alternative complement pathway ,Fast protein liquid chromatography ,Antibody ,Microbiology ,Complement system - Abstract
Complement is a key component of functional immunological assays used to evaluate vaccine-mediated immunity to a range of bacterial and viral pathogens. However, standardization of these assays is complicated due to the availability of a human complement source that lacks existing antibodies acquired either through vaccination or natural circulation of the pathogen of interest. We have developed a method for depleting both IgG and IgM in 200 mL batches from pooled hirudin-derived human plasma by sequential affinity chromatography using a Protein G Sepharose column followed by POROS™ CaptureSelect™ IgM Affinity resin. The production of large IgG- and IgM-depleted batches of human plasma that retains total hemolytic and alternative pathway activities allows for improved assay standardization and comparison of immune responses in large clinical trials.
- Published
- 2021